Metagenome analyses reveal the influence of the inoculant Lactobacillus buchneri CD034 on the microbial community involved in grass ensiling

•First taxonomic profiling of microbial communities involved in a grass ensiling process by applying high-throughput sequencing technologies.•Dominant taxa within ensiling communities in untreated and inoculated silages were identified at a highly resolving level.•Obtained results proved that applic...

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Veröffentlicht in:Journal of biotechnology 2013-09, Vol.167 (3), p.334-343
Hauptverfasser: Eikmeyer, Felix G., Köfinger, Petra, Poschenel, Andrea, Jünemann, Sebastian, Zakrzewski, Martha, Heinl, Stefan, Mayrhuber, Elisabeth, Grabherr, Reingard, Pühler, Alfred, Schwab, Helmut, Schlüter, Andreas
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container_issue 3
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container_title Journal of biotechnology
container_volume 167
creator Eikmeyer, Felix G.
Köfinger, Petra
Poschenel, Andrea
Jünemann, Sebastian
Zakrzewski, Martha
Heinl, Stefan
Mayrhuber, Elisabeth
Grabherr, Reingard
Pühler, Alfred
Schwab, Helmut
Schlüter, Andreas
description •First taxonomic profiling of microbial communities involved in a grass ensiling process by applying high-throughput sequencing technologies.•Dominant taxa within ensiling communities in untreated and inoculated silages were identified at a highly resolving level.•Obtained results proved that application of an inoculant Lactobacillus buchneri strain affects silage characteristics.•Competitiveness of the inoculant L. buchneri strain within the silaging community was shown at the genomic level by performing fragment recruitments of metagenome sequence reads to the complete genome sequence of L. buchneri. Silage is green fodder conserved by lactic acid fermentation performed by epiphytic lactic acid bacteria under anaerobic conditions. To improve the ensiling process and the quality of the resulting silage, starter cultures are added to the fresh forage. A detailed analysis of the microbial community playing a role in grass ensiling has been carried out by high throughput sequencing technologies. Moreover, the influence of the inoculant Lactobacillus buchneri CD034 on the microbial community composition was studied. For this purpose, grass was ensiled untreated or inoculated with L. buchneri CD034. The fresh forage as well as silages after 14 and 58 days of fermentation were characterized physico-chemically. Characteristic silage conditions such as increased titers of lactic acid bacteria and higher concentrations of acetic acid were observed in the inoculated silage in comparison to the untreated samples. Taxonomic community profiles deduced from 16S rDNA amplicon sequences indicated that the relative abundance of Lactococci diminished in the course of fermentations and that the proportion of bacteria belonging to the phyla Proteobacteria and Bacteroidetes increased during the fermentation of untreated silage. In the inoculated silage, members of these phyla were repressed due to an increased abundance of Lactobacilli. In addition, metagenome analyses of silage samples confirmed taxonomic profiles based on 16S rDNA amplicons. Moreover, Lactobacillus plantarum, Lactobacillus brevis and Lactococcus lactis were found to be dominant species within silages as analyzed by means of fragment recruitments of metagenomic sequence reads on complete reference genome sequences. Fragment recruitments also provided clear evidence for the competitiveness of the inoculant strain L. buchneri CD034 during the fermentation of the inoculated silage. The inoculation strain was ab
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Silage is green fodder conserved by lactic acid fermentation performed by epiphytic lactic acid bacteria under anaerobic conditions. To improve the ensiling process and the quality of the resulting silage, starter cultures are added to the fresh forage. A detailed analysis of the microbial community playing a role in grass ensiling has been carried out by high throughput sequencing technologies. Moreover, the influence of the inoculant Lactobacillus buchneri CD034 on the microbial community composition was studied. For this purpose, grass was ensiled untreated or inoculated with L. buchneri CD034. The fresh forage as well as silages after 14 and 58 days of fermentation were characterized physico-chemically. Characteristic silage conditions such as increased titers of lactic acid bacteria and higher concentrations of acetic acid were observed in the inoculated silage in comparison to the untreated samples. Taxonomic community profiles deduced from 16S rDNA amplicon sequences indicated that the relative abundance of Lactococci diminished in the course of fermentations and that the proportion of bacteria belonging to the phyla Proteobacteria and Bacteroidetes increased during the fermentation of untreated silage. In the inoculated silage, members of these phyla were repressed due to an increased abundance of Lactobacilli. In addition, metagenome analyses of silage samples confirmed taxonomic profiles based on 16S rDNA amplicons. Moreover, Lactobacillus plantarum, Lactobacillus brevis and Lactococcus lactis were found to be dominant species within silages as analyzed by means of fragment recruitments of metagenomic sequence reads on complete reference genome sequences. Fragment recruitments also provided clear evidence for the competitiveness of the inoculant strain L. buchneri CD034 during the fermentation of the inoculated silage. The inoculation strain was able to outcompete other community members and also affected physico-chemical characteristics of the silage.</description><identifier>ISSN: 0168-1656</identifier><identifier>EISSN: 1873-4863</identifier><identifier>DOI: 10.1016/j.jbiotec.2013.07.021</identifier><identifier>PMID: 23880441</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>16S rRNA amplicon sequencing ; acetic acid ; anaerobic conditions ; Bacteria - classification ; Bacteria - genetics ; Bacteria - isolation &amp; purification ; biotechnology ; community structure ; DNA, Bacterial - genetics ; Ensiling community ; epiphytes ; Fermentation ; forage ; Fragment recruitment ; grasses ; lactic acid bacteria ; lactic fermentation ; Lactobacillus ; Lactobacillus - classification ; Lactobacillus - genetics ; Lactobacillus - isolation &amp; purification ; Lactobacillus brevis ; Lactobacillus buchneri ; Lactobacillus plantarum ; Lactococcus lactis ; Metagenome ; Metagenome - genetics ; metagenomics ; microbial communities ; Microbial Consortia - genetics ; nucleotide sequences ; physicochemical properties ; Poaceae ; Proteobacteria ; ribosomal DNA ; RNA, Ribosomal, 16S - genetics ; silage ; Silage - microbiology ; silage making ; starter cultures ; Taxonomic profiling</subject><ispartof>Journal of biotechnology, 2013-09, Vol.167 (3), p.334-343</ispartof><rights>2013 Elsevier B.V.</rights><rights>Copyright © 2013 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c389t-263eb64d3cd4a9d6eddb83f0803c6a619eafbbe049a93bc7ebe342685639e29a3</citedby><cites>FETCH-LOGICAL-c389t-263eb64d3cd4a9d6eddb83f0803c6a619eafbbe049a93bc7ebe342685639e29a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.jbiotec.2013.07.021$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,777,781,3537,27905,27906,45976</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23880441$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Eikmeyer, Felix G.</creatorcontrib><creatorcontrib>Köfinger, Petra</creatorcontrib><creatorcontrib>Poschenel, Andrea</creatorcontrib><creatorcontrib>Jünemann, Sebastian</creatorcontrib><creatorcontrib>Zakrzewski, Martha</creatorcontrib><creatorcontrib>Heinl, Stefan</creatorcontrib><creatorcontrib>Mayrhuber, Elisabeth</creatorcontrib><creatorcontrib>Grabherr, Reingard</creatorcontrib><creatorcontrib>Pühler, Alfred</creatorcontrib><creatorcontrib>Schwab, Helmut</creatorcontrib><creatorcontrib>Schlüter, Andreas</creatorcontrib><title>Metagenome analyses reveal the influence of the inoculant Lactobacillus buchneri CD034 on the microbial community involved in grass ensiling</title><title>Journal of biotechnology</title><addtitle>J Biotechnol</addtitle><description>•First taxonomic profiling of microbial communities involved in a grass ensiling process by applying high-throughput sequencing technologies.•Dominant taxa within ensiling communities in untreated and inoculated silages were identified at a highly resolving level.•Obtained results proved that application of an inoculant Lactobacillus buchneri strain affects silage characteristics.•Competitiveness of the inoculant L. buchneri strain within the silaging community was shown at the genomic level by performing fragment recruitments of metagenome sequence reads to the complete genome sequence of L. buchneri. Silage is green fodder conserved by lactic acid fermentation performed by epiphytic lactic acid bacteria under anaerobic conditions. To improve the ensiling process and the quality of the resulting silage, starter cultures are added to the fresh forage. A detailed analysis of the microbial community playing a role in grass ensiling has been carried out by high throughput sequencing technologies. Moreover, the influence of the inoculant Lactobacillus buchneri CD034 on the microbial community composition was studied. For this purpose, grass was ensiled untreated or inoculated with L. buchneri CD034. The fresh forage as well as silages after 14 and 58 days of fermentation were characterized physico-chemically. Characteristic silage conditions such as increased titers of lactic acid bacteria and higher concentrations of acetic acid were observed in the inoculated silage in comparison to the untreated samples. Taxonomic community profiles deduced from 16S rDNA amplicon sequences indicated that the relative abundance of Lactococci diminished in the course of fermentations and that the proportion of bacteria belonging to the phyla Proteobacteria and Bacteroidetes increased during the fermentation of untreated silage. In the inoculated silage, members of these phyla were repressed due to an increased abundance of Lactobacilli. In addition, metagenome analyses of silage samples confirmed taxonomic profiles based on 16S rDNA amplicons. Moreover, Lactobacillus plantarum, Lactobacillus brevis and Lactococcus lactis were found to be dominant species within silages as analyzed by means of fragment recruitments of metagenomic sequence reads on complete reference genome sequences. Fragment recruitments also provided clear evidence for the competitiveness of the inoculant strain L. buchneri CD034 during the fermentation of the inoculated silage. The inoculation strain was able to outcompete other community members and also affected physico-chemical characteristics of the silage.</description><subject>16S rRNA amplicon sequencing</subject><subject>acetic acid</subject><subject>anaerobic conditions</subject><subject>Bacteria - classification</subject><subject>Bacteria - genetics</subject><subject>Bacteria - isolation &amp; purification</subject><subject>biotechnology</subject><subject>community structure</subject><subject>DNA, Bacterial - genetics</subject><subject>Ensiling community</subject><subject>epiphytes</subject><subject>Fermentation</subject><subject>forage</subject><subject>Fragment recruitment</subject><subject>grasses</subject><subject>lactic acid bacteria</subject><subject>lactic fermentation</subject><subject>Lactobacillus</subject><subject>Lactobacillus - classification</subject><subject>Lactobacillus - genetics</subject><subject>Lactobacillus - isolation &amp; purification</subject><subject>Lactobacillus brevis</subject><subject>Lactobacillus buchneri</subject><subject>Lactobacillus plantarum</subject><subject>Lactococcus lactis</subject><subject>Metagenome</subject><subject>Metagenome - genetics</subject><subject>metagenomics</subject><subject>microbial communities</subject><subject>Microbial Consortia - genetics</subject><subject>nucleotide sequences</subject><subject>physicochemical properties</subject><subject>Poaceae</subject><subject>Proteobacteria</subject><subject>ribosomal DNA</subject><subject>RNA, Ribosomal, 16S - genetics</subject><subject>silage</subject><subject>Silage - microbiology</subject><subject>silage making</subject><subject>starter cultures</subject><subject>Taxonomic profiling</subject><issn>0168-1656</issn><issn>1873-4863</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkcGOFCEQhonRuOPqI6gcvXQLDc3QJ2NmdTUZ40H3TICunmVCwwr0JPMOPrSMM-vVExXy1V_114_Qa0paSqh4v2_3xsUCtu0IZS1Zt6SjT9CKyjVruBTsKVpVTjZU9OIKvch5TwjhQ0-fo6uOSUk4pyv0-xsUvYMQZ8A6aH_MkHGCA2iPyz1gFya_QLCA43T5iHbxOhS81bZEo63zfsnYLPY-QHJ4c0MYxzH8pWdnUzSuitk4z0tw5VgVDtEfYKwF3iWdM4aQnXdh9xI9m7TP8OryXqO7z59-br402--3Xzcft41lcihNJxgYwUdmR66HUcA4GskmIgmzQgs6gJ6MgWpWD8zYNRhgvBOyF2yAbtDsGr076z6k-GuBXNTssgVfbUFcsqK8GwTnZJAV7c9o9ZFzgkk9JDfrdFSUqFMQaq8uQahTEIqsVQ2i9r25jFjMDOO_rsfLV-DtGZh0VHqXXFZ3P6pCT6qo6HhXiQ9nAuopDg6SytadohhdAlvUGN1_lvgD8vuohA</recordid><startdate>20130910</startdate><enddate>20130910</enddate><creator>Eikmeyer, Felix G.</creator><creator>Köfinger, Petra</creator><creator>Poschenel, Andrea</creator><creator>Jünemann, Sebastian</creator><creator>Zakrzewski, Martha</creator><creator>Heinl, Stefan</creator><creator>Mayrhuber, Elisabeth</creator><creator>Grabherr, Reingard</creator><creator>Pühler, Alfred</creator><creator>Schwab, Helmut</creator><creator>Schlüter, Andreas</creator><general>Elsevier B.V</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20130910</creationdate><title>Metagenome analyses reveal the influence of the inoculant Lactobacillus buchneri CD034 on the microbial community involved in grass ensiling</title><author>Eikmeyer, Felix G. ; Köfinger, Petra ; Poschenel, Andrea ; Jünemann, Sebastian ; Zakrzewski, Martha ; Heinl, Stefan ; Mayrhuber, Elisabeth ; Grabherr, Reingard ; Pühler, Alfred ; Schwab, Helmut ; Schlüter, Andreas</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c389t-263eb64d3cd4a9d6eddb83f0803c6a619eafbbe049a93bc7ebe342685639e29a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>16S rRNA amplicon sequencing</topic><topic>acetic acid</topic><topic>anaerobic conditions</topic><topic>Bacteria - classification</topic><topic>Bacteria - genetics</topic><topic>Bacteria - isolation &amp; purification</topic><topic>biotechnology</topic><topic>community structure</topic><topic>DNA, Bacterial - genetics</topic><topic>Ensiling community</topic><topic>epiphytes</topic><topic>Fermentation</topic><topic>forage</topic><topic>Fragment recruitment</topic><topic>grasses</topic><topic>lactic acid bacteria</topic><topic>lactic fermentation</topic><topic>Lactobacillus</topic><topic>Lactobacillus - classification</topic><topic>Lactobacillus - genetics</topic><topic>Lactobacillus - isolation &amp; purification</topic><topic>Lactobacillus brevis</topic><topic>Lactobacillus buchneri</topic><topic>Lactobacillus plantarum</topic><topic>Lactococcus lactis</topic><topic>Metagenome</topic><topic>Metagenome - genetics</topic><topic>metagenomics</topic><topic>microbial communities</topic><topic>Microbial Consortia - genetics</topic><topic>nucleotide sequences</topic><topic>physicochemical properties</topic><topic>Poaceae</topic><topic>Proteobacteria</topic><topic>ribosomal DNA</topic><topic>RNA, Ribosomal, 16S - genetics</topic><topic>silage</topic><topic>Silage - microbiology</topic><topic>silage making</topic><topic>starter cultures</topic><topic>Taxonomic profiling</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Eikmeyer, Felix G.</creatorcontrib><creatorcontrib>Köfinger, Petra</creatorcontrib><creatorcontrib>Poschenel, Andrea</creatorcontrib><creatorcontrib>Jünemann, Sebastian</creatorcontrib><creatorcontrib>Zakrzewski, Martha</creatorcontrib><creatorcontrib>Heinl, Stefan</creatorcontrib><creatorcontrib>Mayrhuber, Elisabeth</creatorcontrib><creatorcontrib>Grabherr, Reingard</creatorcontrib><creatorcontrib>Pühler, Alfred</creatorcontrib><creatorcontrib>Schwab, Helmut</creatorcontrib><creatorcontrib>Schlüter, Andreas</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Eikmeyer, Felix G.</au><au>Köfinger, Petra</au><au>Poschenel, Andrea</au><au>Jünemann, Sebastian</au><au>Zakrzewski, Martha</au><au>Heinl, Stefan</au><au>Mayrhuber, Elisabeth</au><au>Grabherr, Reingard</au><au>Pühler, Alfred</au><au>Schwab, Helmut</au><au>Schlüter, Andreas</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Metagenome analyses reveal the influence of the inoculant Lactobacillus buchneri CD034 on the microbial community involved in grass ensiling</atitle><jtitle>Journal of biotechnology</jtitle><addtitle>J Biotechnol</addtitle><date>2013-09-10</date><risdate>2013</risdate><volume>167</volume><issue>3</issue><spage>334</spage><epage>343</epage><pages>334-343</pages><issn>0168-1656</issn><eissn>1873-4863</eissn><abstract>•First taxonomic profiling of microbial communities involved in a grass ensiling process by applying high-throughput sequencing technologies.•Dominant taxa within ensiling communities in untreated and inoculated silages were identified at a highly resolving level.•Obtained results proved that application of an inoculant Lactobacillus buchneri strain affects silage characteristics.•Competitiveness of the inoculant L. buchneri strain within the silaging community was shown at the genomic level by performing fragment recruitments of metagenome sequence reads to the complete genome sequence of L. buchneri. Silage is green fodder conserved by lactic acid fermentation performed by epiphytic lactic acid bacteria under anaerobic conditions. To improve the ensiling process and the quality of the resulting silage, starter cultures are added to the fresh forage. A detailed analysis of the microbial community playing a role in grass ensiling has been carried out by high throughput sequencing technologies. Moreover, the influence of the inoculant Lactobacillus buchneri CD034 on the microbial community composition was studied. For this purpose, grass was ensiled untreated or inoculated with L. buchneri CD034. The fresh forage as well as silages after 14 and 58 days of fermentation were characterized physico-chemically. Characteristic silage conditions such as increased titers of lactic acid bacteria and higher concentrations of acetic acid were observed in the inoculated silage in comparison to the untreated samples. Taxonomic community profiles deduced from 16S rDNA amplicon sequences indicated that the relative abundance of Lactococci diminished in the course of fermentations and that the proportion of bacteria belonging to the phyla Proteobacteria and Bacteroidetes increased during the fermentation of untreated silage. In the inoculated silage, members of these phyla were repressed due to an increased abundance of Lactobacilli. In addition, metagenome analyses of silage samples confirmed taxonomic profiles based on 16S rDNA amplicons. Moreover, Lactobacillus plantarum, Lactobacillus brevis and Lactococcus lactis were found to be dominant species within silages as analyzed by means of fragment recruitments of metagenomic sequence reads on complete reference genome sequences. Fragment recruitments also provided clear evidence for the competitiveness of the inoculant strain L. buchneri CD034 during the fermentation of the inoculated silage. The inoculation strain was able to outcompete other community members and also affected physico-chemical characteristics of the silage.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>23880441</pmid><doi>10.1016/j.jbiotec.2013.07.021</doi><tpages>10</tpages></addata></record>
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subjects 16S rRNA amplicon sequencing
acetic acid
anaerobic conditions
Bacteria - classification
Bacteria - genetics
Bacteria - isolation & purification
biotechnology
community structure
DNA, Bacterial - genetics
Ensiling community
epiphytes
Fermentation
forage
Fragment recruitment
grasses
lactic acid bacteria
lactic fermentation
Lactobacillus
Lactobacillus - classification
Lactobacillus - genetics
Lactobacillus - isolation & purification
Lactobacillus brevis
Lactobacillus buchneri
Lactobacillus plantarum
Lactococcus lactis
Metagenome
Metagenome - genetics
metagenomics
microbial communities
Microbial Consortia - genetics
nucleotide sequences
physicochemical properties
Poaceae
Proteobacteria
ribosomal DNA
RNA, Ribosomal, 16S - genetics
silage
Silage - microbiology
silage making
starter cultures
Taxonomic profiling
title Metagenome analyses reveal the influence of the inoculant Lactobacillus buchneri CD034 on the microbial community involved in grass ensiling
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